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. 2022 Dec 1;11(23):3878. doi: 10.3390/cells11233878

Figure 2.

Figure 2

Normoxic HIF-1α stabilization in HCAECs. (a) Immunoblots representing HIF-1α protein expression in the total cell lysates of HCAECs incubated with medium (control group) or 50 ng/mL each of TNF-α, INF-γ, IL-1β, IL-8, M-CSF, and IGF-I for 6 h, 12 h, 18 h, and 24 h. The immunoblots show two different time courses of HIF-1α stabilization under normoxic conditions. (b) Immunoblots and graphical representation of the fold change of the HIF-1α protein level in the total cell lysates of HCAECs incubated with cell culture medium (control group), 50 ng/mL each of TNF-α, INF-γ, and IL-1β for 12 h or 50 ng/mL each of IL-8, M-CSF, and IGF-I for 18 h. (c) Immunoblots and fold change of the HIF-1α protein level upon treatment with 50 ng/mL of cocktail 1 (TNF-α, INF-γ, and IL-1β) for 12 h or 50 ng/mL of cocktail 2 (IL-8, M-CSF, and IGF-I) for 18 h. (b,c) The data show a significant HIF-1α stabilization upon all treatments. The HIF-1α protein level was normalized to the expression levels of β-actin. The data are represented as the mean ± SEM of three independent experiments (N = 3), * p value ≤ 0.05, ** p value ≤ 0.01, *** p value ≤ 0.001, and **** p value ≤ 0.0001 vs. medium.